Human L1CAM knockout HeLa cell lysate

• WB

Lab

Western blot - Human L1CAM knockout HeLa cell lysate (AB273790)

Lane 1 : Wild-type HeLa cell lysate 20 μg

Lane 2 : L1CAM knockout HeLa cell lysate 20 μg

False colour image of Western blot : Anti-L1CAM antibody [EPR18750] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab208155 was shown to bind specifically to L1CAM. A band was observed at 220 kDa in wild-type HeLa cell lysates with no signal observed at this size in L1CAM knockout cell line ab273836 (knockout cell lysate ab273790). The band observed in the knockout lysate lane below 220 kDa is likely to represent a truncated form of L1CAM. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image wild-type and L1CAM knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-L1CAM antibody [EPR18750] (<a href='/en-us/products/primary-antibodies/l1cam-antibody-epr18750-ab208155'>ab208155</a>) at 1/1000 dilution

All lanes:

Wild-type HeLa cell lysate at 20 µg

Predicted band size: 140 kDa

Observed band size: 220 kDa

false

• WB

Lab

Western blot - Human L1CAM knockout HeLa cell lysate (AB273790)

Lane 1 : Wild-type HeLa cell lysate 20 μg.

Lane 2 : L1CAM knockout HeLa cell lysate 20 μg.

False colour image of Western blot : Anti-L1CAM antibody [EPR23241-224] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab270455 was shown to bind specifically to L1CAM. A band was observed at 220 kDa in wild-type HeLa cell lysates with no signal observed at this size in L1CAM knockout cell line ab273836 (knockout cell lysate ab273790). The band observed in the knockout lysate lane below 220 kDa is likely to represent a truncated form of L1CAM. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image wild-type and L1CAM knockout HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-L1CAM antibody [EPR23241-224] (<a href='/en-us/products/primary-antibodies/l1cam-antibody-epr23241-224-ab270455'>ab270455</a>) at 1/1000 dilution

Lanes 1 and 3:

Wild-type HeLa cell lysate at 20 µg

Lanes 2 and 4:

L1CAM knockout HeLa cell lysate at 20 µg

Lane 5:

A375 cell lysate at 20 µg

Lane 6:

MOLT-4 cell lysate at 20 µg

Predicted band size: 140 kDa

Observed band size: 220 kDa

false

• NGS

Supplier Data

Next Generation Sequencing - Human L1CAM knockout HeLa cell lysate (AB273790)

Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift : 96%